Morphological modifications of knee articular cartilage in bullfrogs (Lithobates catesbeianus) (Anura: Ranidae) during postmetamorphic maturation

The organization of knee articular cartilage of the bullfrog (Lithobates catesbeianus) differs in relation to morphofunctional adaptation in many aspects from similar structures in mammals. Thus, we investigated the structural organization and distribution of the extracellular matrix components in three articular cartilage regions in the distal epiphysis of the femur and proximal epiphysis of the tibia in male bullfrogs at 7, 540 and 1,080 days after metamorphosis. Cartilage thickness and cell density decreased in all regions with age. The basophilia differed among cartilage sites during aging. Calcium deposits were detected in growth cartilage of the femur and tibia in older animals. Immunohistochemical staining for chondroitin-6-sulfate was positive in the pericellular and territorial matrix in all samples. Positive immunostaining for type I collagen was observed in the superficial layer at all ages and in ossification centers of older animals. Reactivity to type II collagen was intense and was found throughout the stroma at all ages. Ultrastructural analysis of the epiphyseal region, in young animals, showed that the cytoplasm of chondrocytes was rich in rough endoplasmic reticulum, Golgi complex and mitochondria. In old animals, were observed a reduction in the size and number of mitochondria, disintegration of rough endoplasmic reticulum, and vacuolization of the Golgi complex. The bullfrog articular cartilage presented structural and organizational changes during aging which may contribute to the functional cartilage deterioration in old animals.     

新疆发现外来入侵物种牛蛙

正牛蛙(Lithobates catesbeianus=Rana catesbeiana)被IUCN列为全球100种最具危害的外来入侵种(Lowe et al. 2000),同时也是我国公布的第一批外来入侵物种名单中惟一的陆栖脊椎动物(王彦平等2006),可通过竞争、捕食和疾病传播等多种方式危害我国两栖类(Wu et al. 2005,Wang et al. 2007,Liu et al. 2013,Liu et al. 2015)。牛蛙目前已成功入侵我国中、东、西南部的广大区域如浙江、安徽、湖南、四川、贵州、云南等地(武正军等2004,     

Landsat-based monitoring of annual wetland change in the Willamette Valley of Oregon,USA from 1972 to 2012

In Oregon’s Willamette Valley, remaining wetlands are at high risk to loss and degradation from agricultural activity and urbanization. With an increased need for fine temporal-scale monitoring of sensitive wetlands, we used annual Landsat MSS and TM/ETM+ images from 1972 to 2012 to manually interpret loss, gain, and type conversion of wetland area in the floodplain of the Willamette River. By creating Tasseled Cap Brightness, Greenness, and Wetness indices for MSS data that visually match TM/ETM+ Tasseled Cap images, we were able to construct a complete and consistent, annual time series and utilize the entire Landsat archive. With an extended time series we were also able to compare annual trends of net change in wetland area before and after the no-net-loss policy established under Section 404 of the Clean Water Act in 1990 using a Theil-Sen Slope estimate analysis. Vegetated wetlands experienced a 314 ha net loss of wetland area and non-vegetated wetlands experienced a 393 ha net gain, indicating higher functioning wetlands were replaced in area by non-vegetated wetland habitats such as agricultural and quarry ponds. The majority of both gain and loss in the study area was attributed to gains and losses of agricultural land. After 1990 policy implementations, the rate of wetland area lost slowed for some wetland categories and reversed into trends of gain in wetland area for others, perhaps representative of the success of increased regulations. Overall accuracy of land use classification through manual interpretation was at 80 %. This accuracy increased to 91.1 % when land use classes were aggregated to either wetland or upland categories, indicating that our methodology was more accurate at distinguishing between general upland and wetland than finer categorical classes.     

Detection of Cryptococcus gattii in Selected Urban Parks of the Willamette Valley, Oregon

Human and animal infections of the fungus Cryptococcus gattii have been recognized in Oregon since 2006. Transmission is primarily via airborne environmental spores and now thought to be locally acquired due to infection in non-migratory animals and humans with no travel history. Previous published efforts to detect C. gattii from tree swabs and soil samples in Oregon have been unsuccessful. This study was conducted to determine the presence of C. gattii in selected urban parks of Oregon cities within the Willamette Valley where both human and animal cases of C. gattii have been diagnosed. Urban parks were sampled due to spatial and temporal overlap of humans, companion animals and wildlife. Two of 64 parks had positive samples for C. gattii. One park had a positive tree and the other park, 60 miles away, had positive bark mulch samples from a walkway. Genotypic subtypes identified included C. gattii VGIIa and VGIIc, both considered highly virulent in murine host models.     

Antipredator Behavior of American Bullfrogs (Lithobates catesbeianus) in a Novel Environment

Invasive species capable of recognizing potential predators may have increased establishment rates in novel environments. Individuals may retain historical predator recognition and invoke innate responses in the presence of taxonomically or ecologically similar predators, generalize antipredator responses, or learn to avoid risky species in novel environments. Invasive amphibians in aquatic environments often use chemical cues to assess predation risk and learn to avoid novel predators via direct experience and/or associated chemical cues. Ontogeny may also influence recognition; experience with predators may need to occur at certain developmental stages for individuals to respond correctly. We tested predator recognition in invasive American bullfrog ( Lithobates catesbeianus) tadpoles that varied in experience with fish predators at the population and individual scale. We found that bullfrog tadpoles responded to a historical predator, largemouth bass ( Micropterus salmoides), only if the population was locally sympatric with largemouth bass. Individuals from a population that did not co‐occur with largemouth bass did not increase refuge use in response to either largemouth bass chemical cues alone or chemical cues with diet cues (largemouth bass fed bullfrog tadpoles). To test whether this behavioral response was generalized across fish predators, we exposed tadpoles to rainbow trout ( Oncorhynchus mykiss) and found that tadpoles could not recognize this novel predator regardless of co‐occurrence with other fish species. These results suggest that environment may be more important for predator recognition than evolutionary history for this invasive species, and individuals do not retain predator recognition or generalize across fish predators.     

Restoration action and species response: oviposition habits of Plebejus icarioides fenderi (Lepidoptera: Lycaenidae) across a restoration chronosequence in the Willamette Valley, Oregon, USA

Habitat restoration is critical to the conservation of rare species. However, restoration efforts often proceed without knowledge of their effects on these species. We investigated the reproductive response of federally endangered Fender’s blue butterfly (Plebejus icarioides fenderi) to prairie restoration in Willamette Valley, Oregon, USA. In 2009 and 2010, we quantified availability of larval host plant, Kincaid’s lupine (Lupinus oreganus), and butterfly oviposition in three restored areas (1–10 years old) and adjacent intact habitat. Oviposition measures in restored areas reached or exceeded intact habitat (0.1 eggs/leaf of host plant and 12 % of time ovipositing) within five years post-restoration. However, none of the restorations provided an equivalent host plant density to intact areas (55 leaves/m²). The different response time of host plants and butterflies to restoration highlights the importance of monitoring both vegetation and oviposition over an ecologically relevant timescale (at least 10 years). For imperiled species, quantifying reproductive response to restoration is critical for adaptive management and successful conservation of the species which restoration efforts are intended to benefit.     

Predicting the potential distribution of the alien invasive American bullfrog (Lithobates catesbeianus) in Brazil

The non-native invasive anuran Lithobates catesbeianus is presently distributed in Brazil, especially in the Atlantic Rainforest biodiversity hotspot. Here, we use a maximum entropy ecological niche modeling algorithm (i) to model the North American native geographic distribution of this species and (ii) to project that model onto the whole of Brazil. After applying a threshold value that balances commission and omission errors, the projection results suggested high probabilities of occurrence mostly in southern and southeastern Brazil. We also present the first report on the species known distribution in Brazil, showing good agreement with model predictions. If the predictive map is interpreted as depicting invasiveness potential of L. catesbeianus, strategies to prevent further invasion in Brazil should be focused especially in the Atlantic Rainforest biodiversity hotspot.     

Diet and Prey Selection of the Invasive American Bullfrog(Lithobates catesbeianus) in Southwestern China

Invasive predators have been widely regarded as one of the principle drivers of the global decline of amphibians, which are among the most threatened vertebrate taxon on Earth. The American bullfrog(Lithobates catesbeianus) is identified as one of the most successful vertebrate invaders and has caused the decline or extinction of some native amphibians in many regions and countries including China. Based on field surveys and stomach content analyses, we examined the diet composition of the invasive bullfrog for the first time in two invaded populations in Yunnan Province, southwestern China, a region of global conservation priority, during the breeding season from 2008 to 2014. Additionally, we conducted the first quantitative study on the prey selection of this global invader among their invaded ranges after controlling for the local anuran assemblage and other aquatic preys in the environment. Our results showed that the range of food items in the stomachs of bullfrogs spanned more than 30 species belonging to ten taxonomic classes. Both of post-metamorphosis individuals and juveniles preyed upon native frogs, independent of the bullfrog's body size and mouth width. Importantly, Jacobs' selection index showed a bullfrog preference for the Yunnan pond frog(Babina pleuraden), one native endemic anuran with population decline, in terms of both food volume and occurrence. We therefore provided direct evidence on the predation impact of the invasive bullfrog on an endemic anuran and urged further efforts to prevent the dispersal of this invader into more fragile habitats to reduce their negative impacts on native amphibians.     

The Absence of Zoonotic Agents in Invasive Bullfrogs (Lithobates catesbeianus) in Belgium and The Netherlands

Exotic invasive bullfrogs (Lithobates catesbeianus) are considered to exert a considerable negative impact on native amphibian communities. This can be due to competition and predation, but they are also a notorious source of the infectious diseases chytridiomycosis and ranavirosis, affecting amphibian populations globally. Little is known regarding their carriage of other microbial agents that might be transferred to humans or other animals. In this study we determined the occurrence of the amphibian pathogens Ranavirus and Batrachochytrium dendrobatidis and of the zoonotic agents Coxiella burnetii, Neospora caninum, Leptospira sp., Toxoplasma gondii, Mycoplasma sp., Campylobacter sp., Salmonella sp. and extended-spectrum beta-lactamase producing Escherichia coli in 164 bullfrogs from three populations in Belgium and The Netherlands. Although B. dendrobatidis was present at a high prevalence of 63%, mean infection loads were low with an average of 10.9 genomic equivalents (SD 35.5), confirming the role of bullfrogs as B. dendrobatidis carriers, but questioning their role as primary reservoirs for B. dendrobatidis transmission to native amphibian communities. All tested samples were negative for the other infectious agents examined. These results suggest a limited role of bullfrogs as carrier of these pathogens.     

The physiology of hibernation in Canadian leopard frogs (Rana pipiens) and bullfrogs (Rana catesbeiana)

Canadian northern leopard frogs (Rana pipiens) and bullfrogs (Rana catesbeiana) were acclimated to 3 degrees C and submerged in anoxic (0-5 mmHg) and normoxic (Po(2) approximately 158 mmHg) water. Periodic measurements of blood Po(2), Pco(2), and pH were made on samples taken anaerobically from subsets of each species. Blood plasma was analyzed for [Na(+)], [K(+)], [Cl(-)], [lactate], [glucose], total calcium, total magnesium, and osmolality. Blood hematocrit was determined, and plasma bicarbonate concentration was calculated. Both species died within 4 d of anoxic submergence. Anoxia intolerance would rule out hibernation in mud, which is anoxic. Both species survived long periods of normoxic submergence (R. pipiens, 125 d; R. catesbeiana, 150 d) with minimal changes in acid-base and ionic status. We conclude that ranid frogs require a hibernaculum where the water has a high enough Po(2) to drive cutaneous diffusion, allowing the frogs to extract enough O(2) to maintain aerobic metabolism, but that an ability to tolerate anoxia for several days may still be ecologically meaningful.     

Host–pathogen dynamics among the invasive American bullfrog (Lithobates catesbeianus) and chytrid fungus (Batrachochytrium dendrobatidis)

Hydrobiologia - The combination of introduced host species and emerging pathogens can result in unanticipated disease dynamics and novel host–pathogen interactions. The American bullfrog...     

舟山群岛变态后牛蛙的食性研究

牛蛙(Ranacatesbeiana)是极其危险的外来入侵种,对当地生物类群的捕食是其主要的危害机制,但目前有关其食性的研究均只局限于一个研究地点。为了全面评估牛蛙对当地生物类群的捕食影响,2005年6月30日至8月11日期间,作者采用洗胃法对舟山群岛8个岛屿(岱山、六横、秀山、佛渡、桃花、虾峙、册子和普陀山)变态后牛蛙的食性进行了研究。共采集到变态后的牛蛙391只,包括牛蛙成体113只和亚成体278只。食性分析结果表明:(1)按食物体积计算,成年牛蛙以十足目、鞘翅目、蜻蜓目、中腹足目、蛙形目和鲤形目为主要食物,亚成体牛蛙以十足目、鞘翅目、鲤形目、蜻蜓目、直翅目、膜翅目、鳞翅目幼虫、中腹足目和蛙形目为主要食物;成体和亚成体牛蛙胃中食物的平均大小均随其体长增加而增大;(2)不同岛屿上的牛蛙在主要食物类型构成上存在差异。这表明牛蛙在不同地方对不同生物类群造成的捕食危害是不一样的,充分反映了其生态危害的广泛性。     

Investigating the dispersal routes used by an invasive amphibian, Lithobates catesbeianus, in human-dominated landscapes

Clarifying how species move across and utilize human-modified landscapes is key to the conservation of declining populations, as well as to the management and control of invasive species. The North American bullfrog (Lithobates catesbeianus) is a globally distributed invasive amphibian that has been implicated in the decline of native amphibians across its invasive range and may also act as a transport vector for a number of deadly amphibian pathogens. Identifying the landscape-level features that facilitate or hinder this species as it moves across an ever-changing landscape is necessary to inform control efforts and limit this species’ impact on already declining amphibian populations. We conducted surveys of 243 wetlands across the Colorado Front Range and used an information-theoretic approach to evaluate the contribution of wetland-specific characteristics and landscape-level factors in determining the detection of bullfrog populations and breeding bullfrog populations. Specifically, our goal was to determine whether features related to overland dispersal or to the connectivity of wetlands were better predictors of bullfrog occurrence. Our results indicated that landscape-level factors that may either hinder or facilitate overland movement, such as topographic complexity and the density of wetlands, were the best predictors of bullfrog occurrence at the scale of our analysis, rather than characteristics relating to the connectivity of wetlands to lotic waterway systems. We suggest that when considering the control or eradication of this species, efforts should be directed at reducing hydroperiod of wetlands and should target regions with a high density of wetlands and/or low topographic relief.     

Diet composition of post-metamorphic bullfrogs (Rana catesbeiana) in the Zhoushan archipelago, Zhejiang Province, China

Bullfrogs (Runa catesbeiana) are listed as one of the 100 worst invasive alien species in the world.They are generalist predators and thus may affect native species through predation.In previous studies,the food contents of bullfrogs were mostly examined at a single site.In the present study,the diet composition of post-metamorphic bullfrogs on eight islands (Daishan,Liuheng,Xiushan,Fodu,Taohua,Xiashi,Cezi,and Putuoshan) in the Zhoushan Archipelago,Zhejiang Province of China,were examined by using the stomach flushing method from June 30 to August 11 in 2005.A total 0f391 individual frogs were measured,including 113 adults and 278 juveniles.The analysis of the stomach contents shows that,for adult bullfrogs,the most important prey (by diet volume) overall were Decapoda,Coleoptera,Odonata,Mesogastropoda,Raniformes,and Cypriniformes.For juvenile bullfrogs,these were Decapoda,Coleoptera,Cypriniformes,Odonata,Orthoptera,Hymenoptera,Lepidoptera larvae,Mesogastropoda,and Raniformes.Moreover,the prey size and diet volume increased with the body size of both adult and juvenile bullfrogs.The diet composition of primary preys of bullfrogs was significantly different among the islands.The results indicate that bullfrogs exert different predatory influences on native fauna at different sites and that bullfrogs are generalist predators with extensive ecological impacts on native fauna.     

Transmission of Batrachochytrium dendrobatidis to wood frogs (Lithobates sylvaticus) via a bullfrog (L. catesbeianus) vector

Chytridiomycosis, an emerging infectious disease caused by the chytrid fungus Batrachochytrium dendrobatidis, threatens anuran populations worldwide. Effects of B. dendrobatidis on frog species are variable. Some species typically develop nonlethal infections and may function as carriers; others typically develop lethal infections that can lead to population declines. Nonlethal infections in the bullfrog (Lithobates catesbeianus) are well-documented. In contrast, recently metamorphosed wood frogs (L. sylvaticus) can die from chytridiomycosis. We conducted an ex-situ experiment between May and July 2010 to determine whether B. dendrobatidis-infected bullfrogs could transmit the fungus to wood frog tadpoles when the two species shared a body of water. We tested for B. dendrobatidis infections with quantitative polymerase chain reactions (qPCR) in a subsample of the wood frog tadpoles and in all metamorphosed wood frogs and compared risk of death of froglets exposed and unexposed to infected bullfrogs. We detected B. dendrobatidis sporadically in subsampled treatment tadpoles (nine of 90, 10%) and frequently in treatment froglets (112 of 113, 99.1%). Pooled risk of froglet death was higher (P<0.001) in treatment enclosures than in control enclosures. Our results indicate that, at the low infection loads bullfrogs tend to carry, swabbing for PCR analyses may underestimate prevalence of B. dendrobatidis in this species. We highlight bullfrog disease screening as a management challenge, especially in light of exotic bullfrog colonies on multiple continents and large-scale global trade in this species. We document the importance of quantifying lethal and sublethal effects of bullfrog vectors on B. dendrobatidis-susceptible species.     

Intracelomic use of tricaine methanesulfonate for anesthesia of bullfrogs (Rana catesbeiana) and leopard frogs (Rana pipiens)

The use of intracelomic injection of dissolved tricaine methanesulfonate (MS-222) as an anesthetic agent in two anuran species was studied. Intracelomic MS-222, at dosages of 100, 250, and 400 mg/kg, rapidly induced tranquilization or anesthesia. Effects were less pronounced or nonexistent at the 50 mg/kg dosage. Depth and duration of anesthesia were dosage related. At the 100, 250, and 400 mg/kg dosages, Rana pipiens attained a greater depth of anesthesia and remained anesthetized for a significantly greater duration than did R. catesbeiana. Dosages of between 250–400 mg/kg reliably induced deep anesthesia without mortality in bullfrogs. Dosages of less than 250 mg/kg are recommended for leopard frogs, since variable mortality was noted with higher dosages. Solubilized tricaine methanesulfonate did not cause gross or histopathological lesions to celomic tissues. Tricaine methanesulfonate injected intracelomically can provide rapid, efficient anesthesia in some anuran species. However, due to the observed intra- and interspecies variation in effect, it should be used cautiously, especially in unfamiliar species. © 1992 Wiley-Liss Inc.     

1200 years of fire and vegetation history in the Willamette Valley, Oregon and Washington, reconstructed using high-resolution macroscopic charcoal and pollen analysis

High-resolution macroscopic charcoal and pollen analyses were used to reconstruct the fire and vegetation history of the Willamette Valley for the last 1200 years. Presented in this paper are three new paleoecological reconstructions from Lake Oswego, Porter Lake, and Warner Lake, Oregon, and portions of previous reconstructions from Battle Ground Lake, Washington, and Beaver Lake, Oregon. The reconstructions show that prior to Euro-American settlement vegetation and fire regimes were influenced by a combination of natural and anthropogenic factors. Battle Ground Lake shows a stronger influence from climate, while Lake Oswego, Beaver Lake, Porter Lake, and Warner Lake were more controlled by human activity. However, human-set fires were also modulated by regional climate variability during the Medieval Climate Anomaly and the Little Ice Age. Fire reconstructions from Battle Ground Lake, Lake Oswego, Beaver Lake, and Porter Lake imply that fires were infrequent in the Willamette Valley 200-300 years prior to Euro-American settlement. The decline of Native American populations due to introduced disease may have led to this reduction in fire activity. The prehistoric record from Warner Lake, however, indicates that fires in the foothills of the Cascade Range were more frequent than on the valley floor, at least until ca. AD 1800. The historic portions of the reconstructions indicate that Euro-American land clearance for agriculture and logging produced the most dramatic shifts in vegetation and fire regimes. All five records indicate that few fires in the Willamette Valley have occurred since ca. AD 1930, and fires today are predominantly grass fires.